Boiling ethidium bromide gel

Vladimir Svetlov svetlov at oncology.wisc.edu
Tue Apr 28 16:27:13 EST 1998


In article <3545f045.25657939 at news.uni-koeln.de>,
a2880709 at smail.uni-koeln.de (Martin Gerken) wrote:

>> Recently I've taken up
> >adding EtBr to the loading buffer instead of the gel - far less EtBr to
> >worry about (in the running buffer etc.), lower background and you can boil
> >the agarose as much as you want.
 > 
 > Unfortunately, EtBr moves in the opposite direction of the DNA...
 > I guess, you have to use far more EtBr as if you put the EtBr directly
 > into the gel.

Unbound EtBr, you mean? But that's good. Let it. Since it's travelling to
the catode from the well (and not from across the entire gel) it's not
gonna affect the brightness of the field at all. Normally you'd have to
wait a while before the bottom part of your gel is cleared from the EtBr.
On the other hand, bound to DNA EtBr travels with it to the anode - these
intercalating dyes don't dissociate that easy - so that yer DNA remains
nice and shiny in the UV. So I don't think that this metaphysical argument
really stands up. 

Regards,
V.

-- 
Vladimir Svetlov, Ph. D.
McArdle Lab for Cancer Research
Dept. Oncology
UW-Madison
1400 University Ave.
Madison, WI 53706



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